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Comparative study of antibody immobilization mediated by lipid and polymer fibers.

ABSTRACT: Antibody immobilization and function retention are important to a variety of applications, including proteomics, drug discovery, diagnostics, and biosensors. The present study investigates antibody immobilization mediated by cholesteryl succinyl silane (CSS) fibers, in comparison to hydrophobic polycaprolactone (PCL) fibers and hydrophilic plasma-treated PCL fibers. When incubated with a model protein, the formation of protein aggregates is observed on hydrophobic PCL fibers but not on the more hydrophobic CSS fibers, indicating that CSS fibers immobilize proteins through mechanisms other than hydrophobic interaction. When exposed to a limited amount of antibody, CSS fibers immobilize more antibodies than plasma-treated PCL fibers and no fewer antibodies than PCL fibers. The function retention of antibodies immobilized on the fibers is analyzed using a cell-capture assay, which shows that the antibody-functionalized CSS fibrous matrices capture 6- or 7-fold more cells than the antibody-functionalized PCL or plasma-treated PCL fibrous matrices, respectively. Data collected from the study show that the lipid fiber-mediated immobilization of antibody not only maintains the advantages of physical immobilization such as easiness and rapidness of operation but also improves function retention.

SUBMITTER: Cohn C

PROVIDER: S-EPMC7067562 | biostudies-literature | 2015 Oct

REPOSITORIES: biostudies-literature

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Comparative study of antibody immobilization mediated by lipid and polymer fibers.

Cohn Celine C Leung Siu Ling SL Zha Zhengbao Z Crosby Jessica J Teng Weibing W Wu Xiaoyi X

Colloids and surfaces. B, Biointerfaces 20150619

Antibody immobilization and function retention are important to a variety of applications, including proteomics, drug discovery, diagnostics, and biosensors. The present study investigates antibody immobilization mediated by cholesteryl succinyl silane (CSS) fibers, in comparison to hydrophobic polycaprolactone (PCL) fibers and hydrophilic plasma-treated PCL fibers. When incubated with a model protein, the formation of protein aggregates is observed on hydrophobic PCL fibers but not on the more ...[more]

PMID: 26141437

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Project description:Immobilization of cell adhesive proteins on the scaffold surface has become a widely reported method that can improve the interaction between scaffold and cells. In this study, three nanofibrous scaffolds obtained by electrospinning of poly(caprolactone) (PCL), poly(L-lactide-co-caprolactone) (PLCL) 70:30, or poly(L-lactide) (PLLA) were subjected to chemical immobilization of gelatin based on aminolysis and glutaraldehyde cross-linking, as well as physisorption of gelatin. Two sets of aminolysis conditions were applied to evaluate the impact of amine group content. Based on the results of the colorimetric bicinchoninic acid (BCA) assay, it was shown that the concentration of gelatin on the surface is higher for the chemical modification and increases with the concentration of free NH2 groups. XPS (X-ray photoelectron spectroscopy) analysis confirmed this outcome. On the basis of XPS results, the thickness of the gelatin layer was estimated to be less than 10 nm. Initially, hydrophobic scaffolds are completely wettable after coating with gelatin, and the time of waterdrop absorption was correlated with the surface concentration of gelatin. In the case of all physically and mildly chemically modified samples, the decrease in stress and strain at break was relatively low, contrary to strongly aminolyzed PLCL and PLLA samples. Incubation testing performed on the PCL samples showed that a chemically immobilized gelatin layer is more stable than a physisorbed one; however, even after 90 days, more than 60% of the initial gelatin concentration was still present on the surface of physically modified samples. Mouse fibroblast L929 cell culture on modified samples indicates a positive effect of both physical and chemical modification on cell morphology. In the case of PCL and PLCL, the best morphology, characterized by stretched filopodia, was observed after stronger chemical modification, while for PLLA, there was no significant difference between modified samples. Results of metabolic activity indicate the better effect of chemical immobilization than of physisorption of gelatin.

Dataset Information

Comparative study of antibody immobilization mediated by lipid and polymer fibers.

Publications

Comparative study of antibody immobilization mediated by lipid and polymer fibers.

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